Blood samples were collected in EDTA-containing tubes, centrifuged, placed in aliquot tubes and stored at −70 to −80°C until analysis. All assays were run after all three studies phases were complete for each subject. For GLP-1, 30 μl of dipeptidyl peptidase IV inhibitor was added to the 4 ml EDTA tube prior to collection. Total GLP-1 assays were performed with Alpco Diagnostics ELISA (43-GPTHU-E01). Insulin concentrations were measured using competitive radioimmunoassay (Millipore). Radioimmunoassays were used to analyze serum leptin (Millipore), serum PYY concentrations (Millipore Cat. #PYYT-66HK), and total serum ghrelin concentrations (Millipore Cat. #GHRT-89HK). All radioimmunoassays were performed with a Perkin Elmer Wallac Gamma counter using Maciel RIA-AID data reduction software. Assays for glucose, triglycerides, and free fatty acids were performed on the Olympus AU400e Chemistry Analyzer (Beckman).
Competitive radioimmunoassay
Manufactured by Merck Group
Sourced in Germany
The Competitive radioimmunoassay is a laboratory technique used to quantify the concentration of a target analyte in a sample. It utilizes the principle of competitive binding between the target analyte and a labeled analogue of the analyte for a limited number of binding sites on an antibody. The concentration of the target analyte in the sample is determined by measuring the amount of labeled analogue that is bound to the antibody.
Automatically generated - may contain errors
Lab products found in correlation
Serum leptin, by Merck Group (2 mentions)
Au400e chemistry analyzer, by Beckman Coulter (2 mentions)
Pyyt 66hk, by Merck Group (2 mentions)
Ezgra 88 k kit, by Merck Group (2 mentions)
Pefabloc sc plus, by Merck Group (2 mentions)
Wallac gamma counter, by PerkinElmer (1 mentions)
Ghrt 89hk, by Merck Group (1 mentions)
4 protocols using competitive radioimmunoassay
1
Biochemical Assays for Metabolic Hormones
2
Multiplex Biomarker Measurement Protocol
3
Ghrelin and Adiponectin Biomarker Analysis
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Blood samples for ghrelin assessment were collected in 2 ml EDTA tubes that were prepared with 100 μl protease inhibitor (Pefabloc® SC Plus, Merck KGaA, Germany). Immediately afterwards, the blood samples were centrifuged for 15 min at 4 • C and 3200g. The blood plasma was then aliquoted and stabilized with HCl. Ghrelin plasma was then frozen at -80 • C until analysis at the Hormone laboratory at Oslo University Hospital. Active (acylated) ghrelin concentrations were determined using the EZGRA-88 K kit (Merck, Germany) in duplicates (total analytical CV at 488 pg/ml 12 %).
Blood samples for adiponectin assessment were collected in 2 ml EDTA tubes that were centrifuged with the same parameters as the ghrelin samples. Afterwards, blood plasma was aliquoted and frozen at -80 • C until analysis at the Hormone Laboratory at Oslo University Hospital. Adiponectin was analysed with competitive radio immuno assay (Merck Millipore, Germany) in duplicates (total analytical CV at 1300 nmol/l 29 %).
Blood samples for adiponectin assessment were collected in 2 ml EDTA tubes that were centrifuged with the same parameters as the ghrelin samples. Afterwards, blood plasma was aliquoted and frozen at -80 • C until analysis at the Hormone Laboratory at Oslo University Hospital. Adiponectin was analysed with competitive radio immuno assay (Merck Millipore, Germany) in duplicates (total analytical CV at 1300 nmol/l 29 %).
4
Blood Biomarker Sampling and Analysis
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Blood samples for ghrelin assessment were collected in 2 ml EDTA tubes that were prepared with 100µl protease inhibitor (Pefabloc® SC Plus, Merck KGaA, Germany). Immediately afterwards, the blood samples were centrifuged for 15 minutes at 4⁰C and 3200 g. The blood plasma was then aliquoted and stabilized with HCl. Ghrelin plasma was then frozen at -80⁰C until analysis at the Hormone laboratory at Oslo University Hospital. Active (acylated) ghrelin concentrations were determined using the EZGRA-88K kit (Merck, Germany) in duplicates (total analytical CV at 488 pg/ml 12%).
Blood samples for adiponectin assessment were collected in 2ml EDTA tubes that were centrifuged with the same parameters as the ghrelin samples. Afterwards, blood plasma was aliquoted and frozen at -80⁰C until analysis at the Hormone Laboratory at Oslo University Hospital. Adiponectin was analysed with competitive radio immuno assay (Merck Millipore, Germany) in duplicates (total analytical CV at 1300 nmol/l 29%).
Blood samples for adiponectin assessment were collected in 2ml EDTA tubes that were centrifuged with the same parameters as the ghrelin samples. Afterwards, blood plasma was aliquoted and frozen at -80⁰C until analysis at the Hormone Laboratory at Oslo University Hospital. Adiponectin was analysed with competitive radio immuno assay (Merck Millipore, Germany) in duplicates (total analytical CV at 1300 nmol/l 29%).
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